Departamento de Ciencia y Tecnología Agroforestal y Genética, ETSIA, Universidad de Castilla-La Mancha, Campus Universitario s/n, E-02071 Albacete, Spain.
J Exp Bot. 2010;61(1):105-19. doi: 10.1093/jxb/erp283.
Crocus sativus is a triploid sterile plant characterized by its long red stigmas, which produce and store significant quantities of carotenoid derivatives formed from the oxidative cleavage of beta-carotene and zeaxanthin. The present study reports on the genomic structures of two lycopene-beta-cyclase genes, CstLcyB1 and CstLcyB2a, and on their transcription patterns in different C. sativus tissues. Phylogenetic analysis showed that both proteins are located in different groups: CstLcyB2a encodes chromoplast-specific lycopene cyclases, with an expression analysis showing strongly in flower stigmas where it activates and boosts beta-carotene accumulation. The CstLcyB1 transcript, however, was present in leaves, tepals, and stigmas at lower levels. In vivo assays in transgenic Arabidopsis demonstrated lycopene beta-cyclase activity of CstLcyB2a. CstLcyB2a is a CstLcyB1 paralogue derived through a gene duplication event, while promoter analysis showed that both genes have diverged in their regulatory sequences after duplication. Furthermore, it was found that the CstLcyB2a gene was absent from Crocus kotschyanus and, although present in C. goulimyi and C. cancellatus, the absence of transcripts suggests that transcriptional regulation of CstLcyB2a is responsible for the low apocarotenoid content in these species.
番红花是一种三倍体不育植物,其特征为长而红色的柱头,这些柱头可产生并储存大量由β-胡萝卜素和玉米黄质氧化裂解形成的类胡萝卜素衍生物。本研究报告了两个番茄红素-β-环化酶基因 CstLcyB1 和 CstLcyB2a 的基因组结构及其在不同番红花组织中的转录模式。系统发育分析表明,这两种蛋白质位于不同的组中:CstLcyB2a 编码质体特异性的番茄红素环化酶,其表达分析表明在花柱头中强烈表达,在那里它激活并促进 β-胡萝卜素的积累。然而,CstLcyB1 转录本在叶片、花瓣和柱头中以较低水平存在。在转基因拟南芥中的体内测定表明 CstLcyB2a 具有番茄红素 β-环化酶活性。CstLcyB2a 是通过基因复制事件衍生的 CstLcyB1 旁系同源物,而启动子分析表明,这两个基因在复制后其调控序列已经分化。此外,还发现 CstLcyB2a 基因在藏红花中不存在,尽管在藏红花和番红花中存在,但没有转录本表明 CstLcyB2a 的转录调控是这些物种中低脱羧类胡萝卜素含量的原因。